JP5947492B2 - Process for producing chiral 8- (3-amino-piperidin-1-yl) -xanthine - Google Patents

Description

  The present invention relates to an improved process for the preparation of chiral 8- (3-aminopiperidin-1-yl) -xanthines, their enantiomers and their physiologically acceptable salts.

  The following general structure

Wherein R ¹ is, for example, an optionally substituted arylmethyl group or an optionally substituted heteroarylmethyl group, R ² is, for example, an alkyl group, and R ³ is, for example, an optionally substituted benzyl group or a linear or branched alkenyl group or alkynyl group)
Of 8- (3-aminopiperidin-1-yl) -xanthine is already known from the international patent applications WO 02/068420, WO04 / 018468, WO04 / 018467, WO2004 / 041820 and WO2004 / 046148 and is useful for these. Compounds with various pharmacological properties (in particular including an inhibitory action on the activity of the enzyme dipeptidyl peptidase IV (DPP-IV)) are described. Therefore, this type of compound is associated with increased DPP-IV activity or is prevented or reduced by a decrease in DPP-IV activity, particularly diabetes mellitus type I or type II, prediabetes, or a decrease in glucose tolerance. Suitable for preventing or treating the resulting disorder or symptom.
WO04 / 018468 discloses a preparation method wherein 8- (3-aminopiperidin-1-yl) -xanthine is prepared by deprotecting the corresponding tert.-butyloxycarbonyl protected derivative of general formula (II) Yes.

  This process results in impurities that were difficult to remove, especially on an industrial scale, due to the protecting groups used. Therefore, the method was unsuitable for the industrial preparation of 8- (3-aminopiperidin-1-yl) -xanthine, especially for drug production with strict requirements on purity. Furthermore, the process has the disadvantage that the preparation of enantiomerically pure precursor 3- (tert.-butyloxycarbonylamino) piperidine is complicated and expensive. However, enantiomerically pure active ingredients are preferred for pharmaceutical applications because of the risk of side effects and are preferred for reducing the dose to a minimum. These situations are disadvantageous for the suitability of the methods known for the industrial preparation of enantiomerically pure 8- (3-aminopiperidin-1-yl) -xanthine.

  In view of the above disadvantages of the known preparation methods, the object of the present invention is to provide enantiomerically pure chirality using starting materials which are easily obtained with high chemical and optical purity without incurring great technical costs and inconveniences. It is to provide a method that allows the preparation of 8- (3-aminopiperidin-1-yl) -xanthine. This new method should also be suitable for industrial scale synthesis and thus commercial use.

This object is achieved by the method of the invention for preparing chiral 8- (3-aminopiperidin-1-yl) -xanthine. In addition to high yield industrial performance, very good chemical and optical purity is a further advantage of the synthesis route of the present invention.
According to the process of the present invention, a suitable xanthine precursor (III) is enantiomerically pure or racemic 3-, at a temperature of 20-160 ° C., preferably 8-140 ° C. in a suitable solvent according to scheme 1. Reacted with (phthalimido) piperidine. The solvent used is, for example, tetrahydrofuran (THF), dioxane, N, N-dimethylformamide (DMF), dimethylacetamide (DMA), N-methyl-2-pyrrolidone (NMP) or dimethyl sulfoxide (DMSO). Also good. NMP is preferably used. Subsequently, the phthalyl protecting group is removed by methods known per se. Possible removal methods are described, for example, in TW Greene “Protecting groups in organic synthesis”, Wiley 1981, p. 265 (eg hydrazine in ethanol).

In the above formula,
X is selected from the group of halogen, for example, fluorine atom, chlorine atom or bromine atom, or sulfonate ester, for example, phenylsulfonyloxy group, p-toluenesulfonyloxy group, methylsulfonyloxy group or trifluoromethylsulfonyloxy group. Leaving group
R ¹ is phenylcarbonylmethyl group, benzyl group, naphthylmethyl group, pyridinylmethyl group, pyrimidinylmethyl group, quinolinylmethyl group, isoquinolinylmethyl group, quinazolinylmethyl group, quinoxalinylmethyl group, naphthyridinylmethyl group Or a phenanthridinylmethyl group, the aromatic or heteroaromatic part being in each case mono- or disubstituted by _Ra , these substituents may be the same or different May be, and
R _a is a hydrogen atom, fluorine atom, chlorine atom or bromine atom or cyano group, methyl group, trifluoromethyl group, ethyl group, phenyl group, methoxy group, difluoromethoxy group, trifluoromethoxy group or ethoxy group, or The two R _a groups may also be a —O—CH ₂ —O— group or a —O—CH ₂ —CH ₂ —O— group if they are attached to adjacent carbon atoms,
R ² is a methyl group, an ethyl group, a propyl group, an isopropyl group, a cyclopropyl group or a phenyl group, and
R ³ is 2-buten-1-yl group, 3-methyl-2-buten-1-yl group, 2-butyn-1-yl group, 2-fluorobenzyl group, 2-chlorobenzyl group, 2-bromobenzyl Group, 2-iodobenzyl group, 2-methylbenzyl group, 2- (trifluoromethyl) benzyl group or 2-cyanobenzyl group.

The method is that X is a chlorine atom or a bromine atom,
R ¹ is phenylcarbonylmethyl group, benzyl group, naphthylmethyl group, pyridinylmethyl group, pyrimidinylmethyl group, quinolinylmethyl group, isoquinolinylmethyl group, quinazolinylmethyl group, quinoxalinylmethyl group or naphthyridinylmethyl group The aromatic or heteroaromatic part is in each case mono- or disubstituted by R _a , these substituents may be the same or different, and
R _a is a hydrogen atom, a fluorine atom or a chlorine atom or a cyano group, a methyl group, an ethyl group, a methoxy group or an ethoxy group,
R ² is a methyl group, an ethyl group, a propyl group, an isopropyl group, a cyclopropyl group or a phenyl group, and
R ³ is 2-buten-1-yl, 3-methyl-2-buten-1-yl, 2-butyn-1-yl, 2-fluorobenzyl, 2-chlorobenzyl, 2-bromobenzyl Preferred for these compounds are the group, 2-iodobenzyl group, 2-methylbenzyl group, 2- (trifluoromethyl) benzyl group or 2-cyanobenzyl group.
The method is that X is a chlorine atom or a bromine atom,
R ¹ is cyanobenzyl group, (cyanopyridinyl) methyl group, quinolinylmethyl group, (methylquinolinyl) methyl group, isoquinolinylmethyl group, (methylisoquinolinyl) methyl group, quinazolinylmethyl group, (methylquinazolinyl) ) Methyl group, quinoxazinylmethyl group, (methylquinoxalinyl) methyl group, (dimethylquinoxalinyl) methyl group or naphthyridinylmethyl group,
R ² is a methyl group, a cyclopropyl group or a phenyl group, and
R ³ is 2-buten-1-yl group, 3-methyl-2-buten-1-yl group, 2-butyn-1-yl group, 2-chlorobenzyl group, 2-bromobenzyl group or 2-cyanobenzyl For these compounds which are radicals, in particular the compound 1-[(4-methylquinazolin-2-yl) methyl] -3-methyl-7- (2-butyn-1-yl) -8- (3- (R ) -Aminopiperidin-1-yl) -xanthine, 1-[(3-methylisoquinolin-1-yl) methyl] -3-methyl-7- (2-butyn-1-yl) -8-((R) -3-aminopiperidin-1-yl) -xanthine and 1-[(3-cyanopiperidin-2-yl) methyl] -3-methyl-7- (2-butyn-1-yl) -8- (3- More preferred for (R) -aminopiperidin-1-yl) -xanthine, where X is bromine.
In each case, it is preferred to use (R) -3- (phthalimido) piperidine as a reagent. The preparation of the compound of formula (III) has already been described in the literature cited above and is carried out by methods known per se.

Furthermore, the present invention provides a method for preparing optically active 3- (phthalimido) piperidine. In this method, 3-aminopyridine is first hydrogenated by methods known per se. The racemic 3-aminopiperidine thus obtained is then converted to the corresponding phthalimide with phthalic anhydride. The (R) enantiomer can be selectively precipitated from a solution of racemic crude phthalimide (IV) with D-tartaric acid. Also, the (S) enantiomer of (IV) can be removed from this salt-precipitated mother liquor in a simple manner by adding L-tartaric acid, without prior removal of excess D-tartaric acid still present in the mother liquor. It is possible to obtain.
This very simple enantiomeric separation of the compound of formula (IV) is surprising to those skilled in the art. The racemic base from the hydrogenation reaction need not be purified in advance for this purpose. The method can be carried out without problems even on an industrial scale.
In addition, the unexpectedly clean reaction between 3-aminopiperidine and phthalic anhydride is surprising in itself. Because, according to the literature (eg US Pat. No. 4,005,208, in particular Example 27), in addition to the desired product, a mixture containing derivatives in which the ring nitrogen atom is acylated is expected. is there.

  The following examples will explain the invention in great detail.

D-tartrate of R enantiomer of 3- (phthalimido) piperidine
a. Hydrogenation

First, 10.00 kg (106.25 mol) of 3-aminopyridine, 500 g of industrial grade activated carbon and 65 liters of acetic acid are charged into a hydrogenation reactor. Add 50 g of Nishimura catalyst (commercially available rhodium / platinum mixed catalyst) as a slurry in 2.5 liters of acetic acid and flush with 2.5 liters of acetic acid. Hydrogenation is carried out at 50 ° C. with a hydrogen pressure of 100 bar until hydrogen absorption ceases, followed by post-hydrogenation at 50 ° C. for 30 minutes. The catalyst and activated carbon are filtered off and washed with 10 liters of acetic acid. The product solution is further reacted without purification.
The reaction also proceeds at a non-harsh pressure.
b. Acylation

15.74 kg (106.25 mol) of phthalic anhydride are initially charged to the reactor and mixed with the filtrate from the hydrogenation. It is flushed with 7.5 liters of acetic acid, the reaction mixture is subsequently heated to reflux and 30% of the acetic acid used during the course is distilled off within 1 hour. The reaction solution is cooled to 90 ° C. The product solution is further reacted without purification.
c. Optical resolution

A solution of 11.16 kg (74.38 mol) of D (−)-tartaric acid in 50 liters of absolute ethanol, heated to 50 ° C., is metered into the acylation reaction solution at 90 ° C. It is flushed with 10 liters of absolute ethanol and stirred for 30 minutes at 90 ° C., during which time the product crystallizes. After cooling to 5 ° C., the product is centrifuged and washed with absolute ethanol. The product solution is further reacted without purification.
d. Heat and reflux the wet crude product in a mixture of 50 liters of acetone and 90 liters of water until a recrystallization solution is formed. Subsequently, the solution is cooled to 5 ° C., during which the product crystallizes. The suspension is stirred for 30 minutes at 5 ° C., the product is centrifuged and finally washed with a mixture of 20 liters of acetone and 10 liters of water. The mixture is dried at 45 ° C. in a drying cabinet under inert conditions.
Yield: 11.7-12.5kg (29-31% of theory)

1-[(4-Methylquinazolin-2-yl) methyl] -3-methyl-7- (2-butyn-1-yl) -8- (3- (R) -aminopiperidin-1-yl) -xanthine
a. 2-Chloromethyl-4-methylquinazoline

First, 10.00 kg (73.98 mol) of 2-aminoacetophenone is charged, and 24.5 liters of 1,4-dioxane is added. The solution, cooled to 10 ° C., is mixed with 16.72 kg (458.68 mol) of hydrogen chloride by gas sealing. The reaction mixture warms to 22-25 ° C. At this temperature, additional hydrogen chloride is gas sealed. From about half of the total gas seal volume, cool the mixture to −10 ° C. and continue the gas seal. Subsequently, the resulting suspension is left at −10 ° C. overnight. A solution of 6.70 kg (88.78 mol) of chloroacetonitrile in 2.5 liters of 1,4-dioxane is added within 1 hour at -10 ° C. Flush the supply container with 2 liters of 1.4-dioxane. Thereafter, the reactor contents are warmed to 6 ° C. and stirred for another 2 hours.
A further reactor is initially charged with a mixture of 122 liters of water and 62.04 kg (775.31 mol) of sodium hydroxide solution (50%) and cooled to 6 ° C. Add the reaction mixture from the first reactor in small portions. The internal temperature is 11 ° C or less. Subsequently, the first reactor is first flushed with 6 liters of 1,4-dioxane and then 6 liters of water. The resulting suspension is stirred at 5 ° C. for a further 30 minutes. The product is centrifuged, washed with 41 liters of water and dried at 35 ° C. in a drying cabinet under inertness.
Yield: 10.5-12.1kg (74-85% of theory)
b. 1-[(4-Methylquinazolin-2-yl) methyl] -3-methyl-7- (2-butyn-1-yl) -8-bromoxanthine

3-methyl-7- (2-butyn-1-yl) -8-bromoxanthine 10.00 kg (33.66 mol), 2-chloromethyl-4-methylquinazoline 7.13 kg (37.02 mol), anhydrous sodium carbonate 3.92 kg (37.02 Mol) and 30 liters of N-methyl-2-pyrrolidone are initially charged to the reactor. The reactor contents are heated to 140 ° C. and stirred at 140 ° C. for 2 hours. After the reaction is complete, the reaction mixture is cooled to 80 ° C. and diluted with 60 liters of 96% ethanol followed by 55 liters of water at 70 ° C. At 60 ° C, weigh 4.04 kg (67.32 mol) of acetic acid and flush with 5 liters of water. The resulting suspension is stirred at 60 ° C. for 30 minutes, then cooled to 23 ° C. and stirred for a further 30 minutes. Subsequently, the product is centrifuged and washed first with a mixture of 20 liters of 96% ethanol and 20 liters of water, then with 40 liters of 96% ethanol and 40 liters of water. Drying is carried out at 45 ° C. in a drying cabinet under inert conditions.
Yield: 11.6-12.6 kg (76-83% of the theoretical value)
c. 1-[(4-Methylquinazolin-2-yl) methyl] -3-methyl-7- (2-butyn-1-yl) -8- (3- (R) -phthalimidopiperidin-1-yl) -Xanthine

1-[(4-Methylquinazolin-2-yl) methyl] -3-methyl-7- (2-butyn-1-yl) -8-bromoxanthine 10.00 kg (22.06 mol), 3- (phthalimido) piperidine D -Charge 12.59 kg (33.09 moles) of tartrate salt and 17.5 liters of N-methyl-2-pyrrolidone to the reactor first. The reactor contents are heated to 140 ° C. After reaching that temperature, 11.41 kg (88.24 mol) of diisopropylethylamine are metered in within 20 minutes. The feed vessel is flushed with 2.5 liters of N-methyl-2-pyrrolidone and the reaction mixture is subsequently stirred at 140 ° C. for 2 hours. After the reaction is complete, the reaction mixture is cooled to 60 ° C. and diluted with 80 liters of methanol. The resulting suspension is stirred at 50 ° C. for 30 minutes, then cooled to 23 ° C. and stirred for a further 30 minutes. Subsequently, the product is centrifuged and washed three times with 20 liters of methanol each time. Drying is carried out at 45 ° C. in a drying cabinet under inert conditions.
Yield: 12.0-12.5kg (90-94% of theoretical value)
d.1-[(4-Methylquinazolin-2-yl) methyl] -3-methyl-7- (2-butyn-1-yl) -8- (3- (R) -aminopiperidin-1-yl) -Xanthine

1-[(4-Methylquinazolin-2-yl) methyl] -3-methyl-7- (2-butyn-1-yl) -8- (3- (R) -phthalimidopiperidin-1-yl) -xanthine 1800 kg (3 mol) is heated to 80-85 ° C. in 18 liters of toluene. Subsequently, 1.815 l (30 mol) of ethanolamine is added to the suspension at 75-80 ° C. In order to complete the reaction, the mixture is stirred at 80-85 ° C. for 2 hours, during which time the solid dissolves in solution. Subsequently, the phases are separated. The ethanolamine phase is washed twice with warm toluene (4 liters each time). The combined toluene phases are washed twice with 8 liters of water each time at 75-80 ° C. From the toluene phase, 22 liters of toluene are distilled off under reduced pressure. 4 liters of tert.-butyl methyl ether are weighed into the suspension obtained at 40-50 ° C. and subsequently cooled to 0-5 ° C. The product is isolated by filtration, washed with tert.-butyl methyl ether and sucked dry. The wet crude material is then heated to reflux with 5 volumes of absolute ethanol and the hot solution is purified by filtration through activated carbon. The filtrate is cooled to 20 ° C. and after crystallization has started, it is diluted with tert.-butyl methyl ether to double the volume. The suspension is cooled to 2 ° C., stirred for a further 2 hours, filtered with suction and dried at 45 ° C. in a vacuum drying cabinet.
Yield: 1174 g (83.2% of the theoretical value)
Alternative method of step d
1-[(4-Methylquinazolin-2-yl) methyl] -3-methyl-7- (2-butyn-1-yl) -8- (3- (R) -phthalimidopiperidin-1-yl) -xanthine 1400 g (2.32 mol) is initially charged in 4.9 liters of tetrahydrofuran and subsequently heated to 55-65 ° C. Subsequently, 350 ml of water and 1433 g (2.32 mol) of ethanolamine are added to the suspension. In order to complete the reaction, the mixture is stirred at 60-63 ° C. for a further 3 hours.

Subsequently, 619 ml of 45% sodium hydroxide solution and 3.85 liters of water are added and the mixture is stirred at 55-65 ° C. for 30 minutes. Then 5.6 liters of toluene are added to the reaction mixture, the mixture is stirred for 15 minutes and subsequently the phases are separated.
The organic phase is washed with 2.8 liters of water at 55-65 ° C. and subsequently removed. From the organic phase, 4.2 liters are distilled off under reduced pressure. Subsequently, 1.4 liters of methylcyclohexane is added at 65-75 ° C., during which the product crystallizes. The suspension is stirred at 15-25 ° C. for 8-16 hours and subsequently cooled to 0-5 ° C. The product is isolated by filtration, washed with 4.2 liters of methylcyclohexane, sucked dry and dried at 35 ° C. under reduced pressure.
The dried crude material (991 g) is then heated to reflux with 5 volumes of methanol, activated carbon is added and the mixture is filtered. The filtrate is reduced to a volume of 1.5 liters by distilling off the methanol. After cooling the filtrate to 45-55 ° C., it is diluted to 4 times volume with tert.-butyl methyl ether. The suspension is cooled to 0-5 ° C., stirred for 2 hours, filtered with suction, washed with tert.-butyl methyl ether and dried at 35 ° C. in a vacuum drying cabinet.
Yield: 899 g (81.9% of theory)

1-[(3-Cyanopyridin-2-yl) methyl] -3-methyl-7- (2-butyn-1-yl) -8- (3- (R) -aminopiperidin-1-yl) -xanthine
a. 3-Cyano-2- (chloromethyl) -pyridine
165.5 g (0.98 mol) of 2-hydroxymethyl-3-pyridinecarboxamide is heated to 90-100 ° C. over 1 hour with 270 ml of phosphorus oxychloride. The reaction mixture is cooled to room temperature and subsequently added dropwise to about 800 ml of water at 50-60 ° C. After hydrolysis of the phosphorus oxychloride, the mixture is neutralized with sodium hydroxide solution with cooling, during which the product precipitates. It is filtered, washed with 300 ml of water and subsequently dried at 35-40 ° C.
Yield: 122.6g (82% of theory)
Alternative method of step a: 3-cyano-2- (chloromethyl) pyridine
20.0 g (131.45 mmol) 2-hydroxymethyl-3-pyridinecarboxamide is suspended in 110 ml acetonitrile and heated to 78 ° C. Within 15 minutes, 60.65 g (395.52 mmol) of phosphorus oxychloride are metered in and the mixture is heated to 81 ° C. for 2 hours. After cooling to 22 ° C., the reaction mixture is stirred at 200 ° C. in 200 ml of water. After adding 100 ml of toluene, the mixture is neutralized with sodium hydroxide solution with cooling. After phase separation, the organic phase is washed with 100 ml of water. The organic phase is removed and the solvent is evaporated under reduced pressure to give an oily residue which crystallizes on standing.
Yield: 16.66g (83% of theory)
b. 1-[(3-Cyanopyridin-2-yl) methyl] -3-methyl-7- (2-butyn-1-yl) -8-bromoxanthine

The reactor was initially charged with 202 g (0.68 mol) of 3-methyl-7- (2-butyn-1-yl) -8-bromoxanthine, 188.5 g (1.36 mol) of anhydrous potassium carbonate and 1.68 liters of N-methyl-2-pyrrolidone. And heat to 70 ° C. Subsequently, 119 g (0.75 mol) of 2-chloromethyl-3-cyanopyridine in 240 ml of N-methyl-2-pyrrolidine (NMP) are added dropwise. The reactor contents are stirred at 70 ° C. for 19 hours. After the reaction is complete, 2.8 liters of water is added to the reaction mixture and it is cooled to 25 ° C. The product is filtered, washed with 2 liters of water and dried at 70 ° C. in a drying cabinet under inertness.
Yield: 257.5g (91% of theory)
c. 1-[(3-Cyanopyridin-2-yl) methyl] -3-methyl-7- (2-butyn-1-yl) -8- (3- (R) -phthalimidopiperidin-1-yl) -Xanthine

1-[(3-Cyanopyridin-2-yl) methyl] -3-methyl-7- (2-butyn-1-yl) -8-bromoxanthine 230 g (0.557 mol), 3- (phthalimido) piperidine D- First, 318 g (0.835 mol) tartrate and 1.15 liters of N-methyl-2-pyrrolidone are charged to the reactor. The reactor contents are heated to 140 ° C. After reaching that temperature, 478 ml (2.78 mol) of diisopropylethylamine are metered in within 20 minutes, and the reaction mixture is subsequently stirred at 140 ° C. for 2 hours. The reaction mixture is subsequently cooled to 75 ° C. and diluted with 720 ml of methanol. Thereafter, 2.7 liters of water are added at 68-60 ° C. and the mixture is cooled to 25 ° C. The product is filtered and washed with 2 liters of water. Drying is carried out at 70 ° C. in a drying cabinet under inert conditions.
The crude product thus obtained is subsequently stirred in 1 liter of boiling methanol, filtered while hot, washed with 200 ml of methanol and subsequently dried at 70 ° C. under inert conditions.
Yield: 275g (88% of theory)
d.1-[(3-Cyanopyridin-2-yl) methyl] -3-methyl-7- (2-butyn-1-yl) -8- (3- (R) -aminopiperidin-1-yl) -Xanthine

1-[(3-Cyanopyridin-2-yl) methyl] -3-methyl-7- (2-butyn-1-yl) -8- (3- (R) -phthalimidopiperidin-1-yl) -xanthine 412.5 g (0.733 mol) is heated to 80 ° C. in 4125 ml of toluene. Subsequently, 445 ml (7.33 mol) of ethanolamine are added to the suspension at 75-80 ° C. In order to complete the reaction, the mixture is stirred at 80-85 ° C. for a further 2 hours, during which time the solid dissolves in solution. Subsequently, the phases are separated. The ethanolamine phase is extracted twice with warm toluene (1 liter each time). The combined toluene phases are washed twice at 75-80 ° C. with 2 liters of water each time. The toluene phase is dried over sodium sulfate, filtered and subsequently reduced to a volume of about 430 ml by distillation under reduced pressure. Subsequently, 1 liter of tert.-butyl methyl ether is metered in at 50-55 ° C. and the mixture is then cooled to 0-5 ° C. The product is isolated by filtration, washed with tert.-butyl methyl ether and dried at 60 ° C. in a drying cabinet.
Yield: 273 g (86% of theory)
Melting point: 188 ± 3 ℃
In the same manner as in Examples 2 and 3, 1-[(3-methylisoquinolin-1-yl) methyl] -3-methyl-7- (2-butyn-1-yl) -8-((R) -3 -Aminopiperidin-1-yl) -xanthine is also prepared.

Claims (4)

A method for producing a medicament for treating type II diabetes mellitus comprising:
i) The following formula (I)

(Where
R ¹ is a (4-methylquinazolin-2-yl) methyl group ,
R ² is a methyl group, and
R ³ is 2-butyn-1-yl group)
A compound of the following synthesis process:
a) the following formula (III)

Wherein X is a leaving group selected from the group of halogen or sulfonic acid esters, and
R ^{1 to} R ³ are defined as above)
Reaction of a compound of (R) -3- (phthalimido) piperidine with
b) the following formula (II)

(Wherein R ^{1 to} R ³ are each defined as above)
Deprotection of the compounds thus obtained
Comprising the steps of: crystallization of the compound of formula (I) from an ethanol solution, and
ii) The method comprising the step of combining the compound of formula (I) with one or more inert carriers and / or diluents. X is Ru Oh chlorine atom or a bromine atom, the process of claim 1. X is Ru Oh bromine atom The method of claim 2 wherein. The method of claim 3 , wherein the solution further comprises t-butyl methyl ether.